BIOZONE SLIDESHOW ENZYMES Enzymes are molecules that act

BIOZONE SLIDESHOW ENZYMES

Enzymes are molecules that act as catalysts to speed up biological reactions. Enzymes are not consumed during the biological reaction. The compound on which an enzyme acts is the substrate. Enzymes can break a single structure into smaller components or join two or more substrate molecules together. Most enzymes are proteins. Many fruits contain enzymes that are used in commercial processes. Pineapple (Ananas comosus, right) contains the enzyme papain which is used in meat tenderization processes and also medically as an anti-inflammatory agent.

Enzyme Examples 3 D molecular structures for the enzymes pepsin (top) and hyaluronidase (bottom). Enzyme Role Pepsin Stomach enzyme used to break protein down into peptides. Works at very acidic p. H (1. 5). Lactase A digestive enzyme that breaks lactose into glucose and galactose. Low levels of lactase can result in lactose intolerance. Topoisomerase A family of enzymes that act on the coiled structure of DNA. They cut the DNA to alter the coiled structure. Hyaluronidase A family of enzymes that break down hyaluronic acid and increase tissue permeability. Often used during eye surgery to administer local anesthetics faster. Zymase A naturally occurring enzyme in yeasts, widely used in the baking industry to ferment sugar into ethanol and carbon dioxide.

Enzymes have a specific region where the substrate binds and where catalysis occurs. This is called the active site. The active site is usually a cleft or pocket at the surface of the enzyme. Substrate modification occurs at the active site. Enzymes are substrate-specific, although specificity varies from enzyme to enzyme: High specificity: The enzyme will only bind with a single type of substrate. Low specificity: The enzyme will bind a range of related substrates, e. g. lipases hydrolyze any fatty acid chain. When a substrate binds to an enzyme’s active site, an enzyme-substrate complex is formed. Space filling model of the yeast enzyme hexokinase. Its active site lies in the groove (arrowed)

Enzyme Active Sites Substrate molecule: Substrate molecules are the chemicals that an enzyme acts on. They are drawn into the cleft of the enzyme. Enzyme molecule: The complexity of the active site is what makes each enzyme so specific (i. e. precise in terms of the substrate it acts on). Active site: The active site contains both binding and catalytic regions. The substrate is drawn to the enzyme’s surface and the substrate molecule(s) are positioned in a way to promote a reaction: either joining two molecules together or splitting up a larger one. This model (above) is an enzyme called Ribonuclease S, that breaks up RNA molecules. It has three active sites (arrowed).

Lock and Key Model The lock and key model of enzyme action, proposed earlier this century, proposed that the substrate was simply drawn into a closely matching cleft on the enzyme molecule. Substrate Enzyme Products Symbolic representation of the lock and key model of enzyme action. 1. A substrate is drawn into the active sites of the enzyme. 2. The substrate shape must be compatible with the enzymes active site in order to fit and be reacted upon. 3. The enzyme modifies the substrate. In this instance the substrate is broken down, releasing two products.

Induced Fit Model More recent studies have revealed that the process is much more likely to involve an induced fit. The enzyme or the reactants (substrate) change their shape slightly. The reactants become bound to enzymes by weak Two substrate molecules are drawn into the cleft of the enzyme. The enzyme changes shape, forcing the substrate molecules to combine. chemical bonds. This binding can weaken bonds within the reactants themselves, allowing the reaction to proceed more readily. The resulting end product is released by the enzyme which returns to its normal shape, ready to undergo more reactions.

Enzymes are catalysts; they make it easier for a reaction to take place. Catalysts speed up reactions by influencing the stability of bonds in the reactants. They may also provide an alternative reaction pathway, thus lowering the activation energy needed for a reaction to take place (see the graph below). Amount of energy stored in the chemicals High Without enzyme: The activation energy required is high. Reactant With enzyme: The activation energy required is lower. High energy Product Low energy Low Start Finish Direction of reaction

Catabolic Reactions Catabolic reactions involve the breakdown of a larger molecules into smaller components, with the release energy (they are exergonic). Enzymes involved in catabolic reactions can cause a single substrate molecule to be drawn into the active site. The substrate is attracted to the enzyme by the “active sites”. The substrate is subjected to stress, which facilitates the breaking of bonds Chemical bonds are broken, causing the substrate molecule to break apart to become two separate molecules. Catabolic reactions include: Enzyme Digestion: Breakdown of large food molecules. Cellular respiration: Oxidative breakdown of fuel molecules such as glucose. The substrate is cleaved and the two products are released to allow the enzyme to work again.

Anabolic Reactions In anabolic reactions, smaller molecules are joined to form larger ones. The substrate is attracted to the enzyme by the “active sites”. These reactions are endergonic; they require the input of energy. The substrate is subjected to stress, which will aid the formation of bonds. Enzymes involved in anabolic reactions can cause two substrate molecules to be drawn into the active site. New chemical bonds are formed resulting in the formation of a single molecule. Enzyme Examples include: Protein synthesis: Build up of polypeptides from peptide units. Cellular respiration: Oxidative breakdown of fuel molecules such as glucose. The two substrate molecules form a single product, which is released, freeing the enzymes to work again.

Effect of Temperature Enzyme activity increases with Optimum temperature for the enzyme Rate of reaction Enzymes often have a narrow range of conditions under which they operate properly. For most plant and animal enzymes, there is little activity at low temperatures. Rapid denaturation at high temperatures Too cold for the enzyme to operate temperature, until the temperature is too high for the enzyme to function. (See diagram right). At this point, enzyme denaturation occurs and the enzyme can no longer function. Temperature (°C)

Effect of p. H Enzymes can be affected by p. H. Extremes of p. H (very acid or alkaline) away enzyme denaturation. Enzymes are found in very diverse p. H conditions, so they must be suited to perform in these specialist environments. Pepsin is a stomach enzyme and has an optimal working p. H of 1. 5, which is suited for the very acidic conditions of the stomach. Urease breaks down urea and has an optimal p. H of near neutral. See diagram right. Enzyme activity from the enzyme optimum can result in Trypsin Urease Pepsin 1 2 3 4 5 Acid 6 7 8 9 10 Alkaline p. H Enzymes often work over a range of p. H values, but all enzymes have an optimum p. H where their activity rate is fastest.

Factors Affecting Enzyme Reaction Rates Effect of Substrate Concentration Rate of reaction Effect of Enzyme Concentration Enzyme concentration Concentration of substrate Rate of reaction continues to increase with an increase in enzyme concentration. Rate of reaction increases and then plateaus with increasing substrate concentration. This relationship assumes non-limiting amounts of substrate and cofactors. This relationship assumes a fixed amount of enzyme.

Enzyme Cofactors Some enzymes require cofactors to be active. Cofactors are a nonprotein component of an enzyme. Cofactors can be: organic molecules (coenzymes). Active site Enzyme is protein only Example: lysozyme Enzyme Active site Enzyme Prostheti c group Enzyme + prosthetic group Example: flavoprotein + FAD inorganic ions (e. g. Ca 2+, Zn 2+). Cofactors may be: Permanently attached, in which case they are called prosthetic groups. Temporarily attached coenzymes, which detach after a reaction, and may participate with another enzyme in other reactions. Active site Coenzyme Enzyme + coenzyme Example: dehydrogenases + NAD

Enzyme Inhibitors Reversible inhibitors are used to control enzyme activity. There is often an interaction between the substrate or end product and the enzymes controlling the reaction. Irreversible inhibitors bind tightly and permanently to the enzymes destroying their catalytic activity. Irreversible inhibitors usually covalently modify an enzyme. Many drug molecules are enzyme inhibitors. Native arsenic Mercury Photo: US EPA Enzymes can be deactivated by enzyme inhibitors. There are two types of enzyme inhibitors: Some heavy metals (above) are examples of poisons which act as irreversible enzyme inhibitors.

Irreversible Enzyme Inhibitors Substrate Some heavy metals, such as cadmium (Cd), arsenic (As), and lead (Pb) act as irreversible enzyme inhibitors. The lipothiamide pyrophosphatase enzyme with substrate bound to its active site. Enzyme They bind strongly to the sulphydryl (SH) groups of the protein, destroying its catalytic activity. Most heavy metals, e. g. arsenic, act as non-competitive inhibitors. Mercury (Hg) is an exception. It acts as a competitive inhibitor, binding directly to a sulphydryl group in the active site of the papain enzyme. Heavy metals are retained in the body, and lost slowly. As Arsenic binds to the enzyme and causes its shape to change, preventing the substrate from binding to the active site. Poisons, such as arsenic (As), act as an irreversible enzyme inhibitor. It binds to the lipothiamide pyrophosphatase enzyme altering its shape so the substrate cannot bind.

Reversible Inhibitors Reversible inhibitors are used to control enzyme activity. There is often an interaction between the substrate or end product and the enzymes controlling the reaction. Buildup of the end product or a lack of substrate may deactivate the enzyme. Competitive inhibition involves competition for the active site. Noncompetitive inhibitors work either to slow down the rate of reaction, or block the active site altogether and prevent its functioning (allosteric inhibition). Substrate Competitive inhibitor blocks the active site. The substrate cannot bind. S The substrate can still bind to the active site but the rate of reaction is lowered. S Enzyme S S Enzyme Noncompetitive inhibitor No inhibition The substrate cannot bind to the active site because the active site is distorted. Competitive inhibition Noncompetitive inhibition Enzyme Noncompetitive inhibitor Allosteric enzyme inhibitor